Small molecule inhibitors of rotamase enzyme activity

ABSTRACT

This invention relates to neurotrophic N-glyoxyl-prolyl ester compounds having an affinity for FKBP-type immunophilins, their preparation and use as inhibitors of the enzyme activity associated with immunophilin proteins, and particularly inhibitors of peptidyl-prolyl isomerase or rotamase enzyme activity.

RELATED APPLICATION

This application is a Continuation-in-Part Application of U.S. patentapplication Ser. No. 08/479,436, filed Jun. 7, 1995, now U.S. Pat. No.5,614,547.

BACKGROUND OF THE INVENTION

1. Field of the Invention

This invention relates to neurotrophic compounds having an affinity forFKBP-type immunophilins, their preparation and use as inhibitors of theenzyme activity associated with immunophilin proteins, and particularlyinhibitors of peptidyl-prolyl isomerase or rotamase enzyme activity.

2. Description of the Prior Art

The term immunophilin refers to a number of proteins that serve asreceptors for the principal immunosuppressant drugs, cyclosporin A(CsA), FK506, and rapamycin. Known classes of immunophilins arecyclophilins, and FK506 binding proteins, such as FKBP. Cyclosporin Abinds to cyclophilin while FK506 and rapamycin bind to FKBP. Theseimmunophilin-drug complexes interface with a variety of intracellularsignal transduction systems, especially in the immune system and thenervous system.

Immunophilins are known to have peptidyl-prolyl isomerase (PPIase) orrotamase enzyme activity. It has been determined that rotamase activityhas a role in the catalyzation of the interconversion of the cis andtrans isomer of immunophilin proteins.

Immunophilins were originally discovered and studied in immune tissue.It was initially postulated by those skilled in the art that inhibitionof the immunophilins rotamase activity leads to the inhibition of T-cellproliferation, thereby causing the immunosuppressive action exhibited byimmunosuppressive drugs such as cyclosporin A, FK506, and rapamycin.Further study has shown that the inhibition of rotamase activity, in andof itself, is not sufficient for immunosuppressant activity. Schreiberet al., Science, 1990 vol. 250 pp. 556-559. It has been shown that theimmunophilin-drug complexes interact with ternary protein targets astheir mode of action. Schreiber et al., Cell, 1991, vol. 66, pp.807-815. In the case of FKBP-FK506 and FKBP-CsA, the drug-immunophilincomplexes bind to the enzyme calcineurin, inhibitory T-cell receptorsignalling leading to T-cell proliferation. Similarly, the complex ofrapamycin and FKBP interacts with the RAFT1/FRAP protein and inhibitssignalling from the IL-2 receptor.

Immunophilins have been found to be present at high concentrations inthe central nervous system. Immunophilins are enriched 10-50 times morein the central nervous system than in the immune system. Within neuraltissues, immunophilins appear to influence neuronal process extension,nitric oxide synthesis, and neurotransmitter release.

It has been found that picomolar concentrations of an immunosuppressantsuch as FK506 and rapamycin stimulate neurite out growth in PC12 cellsand sensory nervous, namely dorsal root ganglion cells (DRGs). Lyons etal., Proc. of Natl. Acad. Sci., 1994 vol. 91, pp. 3191-3195. In wholeanimal experiments, FK506 has been shown to stimulate nerve regenerationfollowing facial nerve injury and results in functional recovery inanimals with sciatic nerve lesions.

Surprisingly, it has been found that drugs with a high affinity for FKBPare potent rotamase inhibitors causing a neurotrophic effect. Lyons etal. These findings suggest the use of immunosuppressants in treatingvarious peripheral neuropathies and enhancing neuronal regrowth in thecentral nervous system (CNS). Studies have demonstrated thatneurodegenerative disorders such as Alzheimer's disease, Parkinson'sdisease, and amyotrophic lateral sclerosis (ALS) may occur due to theloss, or decreased availability, of a neurotrophic substance specificfor a particular population of neurons affected in the disorder.

Several neurotrophic factors effecting specific neuronal populations inthe central nervous system have been identified. For example, it hasbeen hypothesized that Alzheimer's disease results from a decrease orloss of nerve growth factor (NGF). It has thus been proposed to treatAlzheimer's patients with exogenous nerve growth factor or otherneurotrophic proteins such as brain derived nerve factor (BDNF), glialderived nerve factor, ciliary neurotrophic factor, and neurotropin-3 toincrease the survival of degenerating neuronal populations.

Clinical application of these proteins in various neurological diseasestates is hampered by difficulties in the delivery and bioavailabilityof large proteins to nervous system targets. By contrast,immunosuppressant drugs with neurotrophic activity are relatively smalland display excellent bioavailability and specificity. However, whenadministered chronically, immunosuppressants exhibit a number ofpotentially serious side effects including nephrotoxicity, such asimpairment of glomerular filtration and irreversible interstitialfibrosis (Kopp et al., 1991, J. Am. Soc. Nephrol. 1:162); neurologicaldeficits, such as involuntary tremors, or non-specific cerebral anginasuch as non-localized headaches (De Groen et al., 1987, N. Engl. J. Med.317:861); and vascular hypertension with complications resultingtherefrom (Kahan et al., 1989 N. Engl. J. Med. 321: 1725).

In order to prevent the side effects associated with use of theimmunosuppressant compounds, the present invention providesnon-immunosuppressive compounds containing small molecule FKBP rotamaseinhibitors for promoting neuronal growth and regeneration in variousneuropathological situations where neuronal repair can be facilitatedincluding peripheral nerve damage by physical injury or disease statesuch as diabetes, physical damage to the central nervous system (spinalcord and brain) brain damage associated with stroke, and for thetreatment of neurological disorders relating to neurodegeneration,including Parkinson's disease, Alzheimer's disease, and amyotrophiclateral sclerosis.

SUMMARY OF THE INVENTION

The present invention relates to a novel class of neurotrophic compoundshaving an affinity for FKBP-type immunophilins. Once bound to thisprotein the neurotrophic compounds are potent inhibitors of the enzymeactivity associated with immunophilin proteins and particularly rotamaseenzyme activity, thereby stimulating neuronal regeneration andoutgrowth. A key feature of the compounds of the present invention isthat they do not exert any significant immunosuppressive activity inaddition to their neurotrophic activity.

A preferred embodiment of this invention is a neurotrophic compound ofthe formula: ##STR1## where R₁ is selected from the group consisting ofa C₁ -C₉ straight or branched chain alkyl or alkenyl group optionallysubstituted with C₃ -C₈ cycloalkyl, C₃ or C₅ cycloalkyl, C₅ -C₇cycloalkenyl, Ar₁, where said alkyl, alkenyl, cycloalkyl or cycloalkenylgroups may be optionally substituted with C₁ -C₄ alkyl, C₁ -C₄ alkenyl,or hydroxy, where Ar₁ is selected from the group consisting of1-napthyl, 2-napthyl, 2-indolyl, 3-indolyl, 2-furyl, 3-furyl,2-thiazolyl, 2-thienyl, 3-thienyl, 2-,3-, 4-pyridyl, and phenyl, havingone to three substituents which are independently selected from thegroup consisting of hydrogen, halo, hydroxyl, nitro, trifluoromethyl, C₁-C₆ straight or branched alkyl or alkenyl, C₁ -C₄ alkoxy or C₁ -C₄alkenyloxy, phenoxy, benzyloxy, and amino;

X is selected from the group consisting of oxygen, sulfur, methylene(CH₂) , or H₂ ;

Y is selected from the group consisting of oxygen or NR₂, where R₂ ishydrogen or C₁ -C₆ alkyl; and

Z is selected from the group consisting of C₂ -C₆ straight or branchedchain alkyl or alkenyl,

wherein the alkyl chain is substituted in one or more positions with Ar₁as defined above, C₃ -C₈ cycloalkyl, cycloalkyl connected by a C₁ -C₆straight or unbranched alkyl or alkenyl chain, and Ar₂ where Ar₂ isselected from the group consisting of 2-indolyl, 3-indolyl, 2-furyl,3-furyl, 2- thiazolyl, 2-thienyl, 3-thienyl, 2-, 3-, or 4-pyridyl, andphenyl, having one to three substituents which are independentlyselected from the group consisting of hydrogen, halo, hydroxyl, nitro,trifluoromethyl, C₁ -C₆ straight or branched alkyl or alkenyl, C₁ -C₄alkoxy or C₁ -C₄ alkenyloxy, phenoxy, benzyloxy, and amino;

Z may also be the fragment: ##STR2## where R₃ is selected from the groupconsisting of straight or branched alkyl C₁ -C₈ optionally substitutedwith C₃ -C₈ cycloalkyl, or Ar₁ as defined above, and unsubstituted Ar₁ ;

X₂ is O or NR₅, where R₅ is selected from the group consisting ofhydrogen, C₁ -C₆ straight or branched alkyl and alkenyl;

R₄ is selected from the group consisting of phenyl, benzyl, C₁ -C₅straight or branched alkyl or alkenyl, and C₁ -C₅ straight or branchedalkyl or alkenyl substituted with phenyl; or pharmaceutically acceptablesalts or hydrates thereof.

Another preferred embodiment of this invention is a neurotrophiccompound of the formula: ##STR3## where R₁ is a C₁ -C₉ straight orbranched chain alkyl or alkenyl group optionally substituted with C₃ -C₈cycloalkyl, C₃ or C₅ cycloalkyl, C₅ -C₇ cycloalkenyl, or Ar₁, where saidalkyl, alkenyl, cycloalkyl or cycloalkenyl groups may be optionallysubstituted with C₁ -C₄ alkyl, C₁ -C₄ alkenyl, or hydroxy, and where Ar₁is selected from the group consisting of 1-napthyl, 2-napthyl,2-indolyl, 3-indolyl, 2-furyl, 3-furyl, 2-thiazolyl, 2-thienyl,3-thienyl, 2-,3-, or 4-pyridyl, or phenyl, having one to threesubstituents which are independently selected from the group consistingof hydrogen, halo, hydroxyl, nitro, trifluoromethyl, C₁ -C₆ straight orbranched alkyl or alkenyl, C₁ -C₄ alkoxy or C₁ -C₄ alkenyloxy, phenoxy,benzyloxy, and amino;

Z is a C₂ -C₆ straight or branched chain alkyl or alkenyl, wherein thealkyl chain is substituted in one or more positions with Ar₁ as definedabove, C₃ -C₈ cycloalkyl, cycloalkyl connected by a C₁ -C₆ straight orunbranched alkyl or alkenyl chain, or Ar₂ where Ar₂ is selected from thegroup consisting of 2-indolyl, 3-indolyl, 2-furyl, 3-furyl, 2-thiazolyl, 2-thienyl, 3-thienyl, 2-, 3-, or 4-pyridyl, or phenyl, havingone to three substituents which are independently selected from thegroup consisting of hydrogen, halo, hydroxyl, nitro, trifluoromethyl, C₁-C₆ straight or branched alkyl or alkenyl, C₁ -C₄ alkoxy or C₁ -C₄alkenyloxy, phenoxy, benzyloxy, and amino; or pharmaceuticallyacceptable salts or hydrates thereof.

Another preferred embodiment of the invention is a neurotrophic compoundhaving an affinity for FKBP-type immunophilins which inhibit therotamase activity of the immunophilin.

Another preferred embodiment of the present invention is a method fortreating a neurological disorder in an animal comprising administering atherapeutically effective amount of a compound having an affinity forFKBP-type immunophilins which inhibits the rotamase activity of theimmunophilin.

Another preferred embodiment of the invention is a method of promotingneuronal regeneration and growth in mammals, comprising administering toa mammal an effective amount of a neurotrophic compound having anaffinity for FKBP-type immunophilins which inhibits the rotamaseactivity of the immunophilin.

Yet another preferred embodiment of the invention is a method ofpreventing neurodegeneration in an animal comprising administering to ananimal an effective amount of a neurotrophic compound having an affinityfor FKBP-type immunophilins which inhibits rotamase activity of theimmunophilin.

Another preferred embodiment is a neurotrophic N-glyoxyl prolyl estercompound of the formula: ##STR4## where R₁ is a C₁ -C₅ straight orbranched chain alkyl or alkenyl group optionally substituted with C₃ toC₆ cycloalkyl, or Ar₁, where Ar₁ is selected from the group consistingof 2-furyl, 2-thienyl, or phenyl;

X is selected from the group consisting of oxygen and sulfur;

Y is oxygen; and

Z is a straight or branched chain alkyl or alkenyl, wherein the alkylchain is substituted in one or more positions with Ar₁ as defined above,C₃ -C₆ cycloalkyl, Ar₂ where Ar₂ is selected from the group consistingof 2-, 3-, or 4-pyridyl, or phenyl, having one to three substituentswhich are independently selected from the group consisting of hydrogenand C₁ -C₄ alkoxy.

Particularly preferred neurotrophic N-glyoxyl prolyl ester compoundsaccording to the above formula are selected from the group consistingof:

3-(2,5-dimethoxyphenyl)-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,

3-(2,5-dimethoxyphenyl)-1-prop-2-(E)-enyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,

2-(3,4,5-trimethoxyphenyl)-1-ethyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,

3-(3-Pyridyl)-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,

3-(2-Pyridyl)-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,

3-(4-Pyridyl)-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,

3-phenyl-1-propyl(2S)-1-(2-tert-butyl-1,2-dioxoethyl)-2-pyrrolidinecarboxylate,

3-phenyl-1-propyl(2S)-1-(2-cyclohexylethyl-1,2-dioxoethyl)-2-pyrrolidinecarboxylate,

3-(3-pyridyl)-1-propyl(2S)-1-(2-cyclohexylethyl-1,2-dioxoethyl)-2-pyrrolidinecarboxylate,

3-(3-pyridyl)-1-propyl(2S)-1-(2-tert-butyl-1,2-dioxoethyl)-2-pyrrolidinecarboxylate,

3,3-diphenyl-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,

3-(3-pyridyl)-1-propyl(2S)-1-(2-cyclohexyl-1,2-dioxoethyl)-2-pyrrolidinecarboxylate,

3-(3-Pyridyl)-1-propyl (2S)-N-( 2-thienyl!glyoxyl)pyrrolidinecarboxylate,

3,3-Diphenyl-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxobutyl)-2-pyrrolidinecarboxylate,

3,3-Diphenyl-1-propyl (2S)-1-cyclohexylglyoxyl-2-pyrrolidinecarboxylate,and

3,3-Diphenyl-1-propyl(2S)-1-(2-thienyl)glyoxyl-2-pyrrolidinecarboxylate.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a photomicrograph of chick dorsal root ganglia treated withvarious concentrations of Example 17 as indicated. FIG. 1 shows thatExample 17 of the present invention potently promotes neurite outgrowthin sensory neuronal cultures. Explant cultures isolated form embryonicday 9-10 chick dorsal root ganglia were treated with variousconcentrations of Example 17 as indicated. Forty-eight hours later, thenumber of neurite with a length greater than one DRG explant wasquantitated. The number of neurites expressed in untreated DRG's wassubtracted form the neurite number of Example 17-treated samples toyield Example 17-dependent specific neurite outgrowth. Micrographs ofExample 17 treated DRG's, as well as quantitative dose-dependent neuriteoutgrowth elicited by Example 17 are presented.

FIG. 2 is a graph showing quantitation of neurite outgrowth in chickdorsal root ganglia treated with various concentrations of Example 17 asindicated. FIG. 2 shows that Example 17 of the present inventionpotently promotes neurite outgrowth in sensory neuronal cultures.Explant cultures isolated form embryonic day 9-10 chick dorsal rootganglia were treated with various concentrations of Example 17 asindicated. Forty-eight hours later, the number of neurite with a lengthgreater than one DRG explant was quantitated. The number of neuritesexpressed in untreated DRG's was subtracted form the neurite number ofExample 17-treated samples to yield Example 17-dependent specificneurite outgrowth. Quantitative dose-dependent neurite outgrowthelicited by Example 17 is presented.

FIG. 3 is a photomicrograph of rat sciatic nerve sections. FIG. 3 showsthat Example 1 of the present invention promotes neuronal regenerationfollowing sciatic nerve lesions. Sciatic nerves of 150 g maleSprague-Dawley rats were crushed at the level of the hips. Example 1 (30mg/kg s.c.), Inactive (30 mg/kg s.c.) or intralipid vehicle wasadministered once daily for the next 21 days. Animals were sacrificed,sciatic nerves removed and nerve segments 2 mm distal to the crush sitewere sectioned and stained with Holmes silver stain (to assess axonnumber) and Luxol fast blue (to assess remyelination). The micrographsshow sciatic nerve sections of sham operated rats, vehicle-treatedlesioned animals, Example 1 and Inactive treated at 630× magnification,four animals per group.

FIG. 4 is a graph of ³ H! -CFT binding per μg of Striatal MembraneProtein. FIG. 4 shows that neuroimmunophilin ligands of the presentinvention promote recovery of dopamine neurons following MPTP treatmentof mice. CD1 mice (25 g) were treated daily with 30 mg/kg MPTP (i.p.)for 5 days. The animals were also treated daily with intralipid vehicle,Example 1 (100 mg/kg s.c.) or Example 17 (40, 20, 10 mg/kg s.c., asindicated) concurrently with the MPTP and continued for an additional 5days. After eighteen days, the mice were sacrificed, striata from 5animals per group were pooled and processed into a washed membranepreparation. Binding of 3H!-CFT to these striated membrane preparationsof various groups was quantitated to determine dopamine transporterlevels on viable nerve terminals. Binding in the presence of 10 μMunlabelled CFT provided on estimate of nonspecific binding, which wassubtracted from the total binding to quantitative specific 3H!-CFTbound. Binding was normalized to the protein content of the striatalmembranes from each experimental group. Coronal and saggital brainsections from MPTP and drug treated animals were stained withanti-tyrosine hydroxylase (TH) Ig to quantitate striatal, medialforebrain bundle axonal and nigral levels of TH, which is indicative offunctional dopaminergic neurons.

FIG. 5 is a bar graph of ³ H! -CFT plotted for 200 μg of membraneprotein. FIG. 5 shows that neuroimmunophilin ligands of the presentinvention promote recovery of dopamine neurons following MPTP treatmentof mice in accordance with the procedure described in FIG. 4.

FIG. 6 is a photomicrograph, at 630× magnification, of coronal andsaggital brain sections. FIG. 6 shows brain sections from MPTP and drugtreated animals stained with anti-tyrosine hydroxylase (TH) Ig toquantitate striatal levels of TH, which is indicative of functionaldopaminergic neurons.

FIG. 7 is a photomicrograph, at 50× magnification, of coronal andsaggital brain sections. FIG. 7 shows brain sections from MPTP and drugtreated animals stained with anti-tyrosine hydroxylase (TH) Ig toquantitate nigral levels of TH, which is indicative of functionaldopaminergic neurons.

FIG. 8 is a photomicrograph, at 400× magnification, of coronal andsaggital brain sections. FIG. 8 shows brain sections from MPTP and drugtreated animals stained with anti-tyrosine hydroxylase (TH) Ig toquantitate medial forebrain bundle axonal levels of TH, which isindicative of functional dopaminergic neurons.

DETAILED DESCRIPTION OF THE INVENTION

The novel neurotrophic compounds of this invention are relatively smallmolecules in relation to other known compounds which bind to FKBP-typeimmunophilins, such as rapamycin, FK506, and cyclosporin.

The neurotrophic compounds of this invention have an affinity for theFK506 binding proteins such as FKBP-12. When the neurotrophic compoundsof the invention are bound to the FKBP, they have been found tounexpectedly inhibit the prolyl- peptidyl cis-trans isomerase activity,or rotamase activity of the binding protein and stimulate neuritegrowth, while not exhibiting an immunosuppressant effect.

More particularly, this invention relates to a novel class ofneurotrophic compounds represented by the formula: ##STR5## where R₁ isa C₁ -C₉ straight or branched chain alkyl or alkenyl group optionallysubstituted with C₃ -C₈ cycloalkyl, C₃ or C₅ cycloalkyl, C₅ -C₇cycloalkenyl, or Ar₁, where said alkyl, alkenyl, cycloalkyl orcycloalkenyl groups may be optionally substituted with C₁ -C₄ alkyl,C_(1-C) ₄ alkenyl, or hydroxy, and where Ar₁ is selected from the groupconsisting of 1-napthyl, 2-napthyl, 2-indolyl, 3-indolyl, 2-furyl,3-furyl, 2- thiazolyl, 2-thienyl, 3-thienyl, 2-, 3-, or 4-pyridyl, orphenyl, having one to three substituents which are independentlyselected from the group consisting of hydrogen, halo, hydroxyl, nitro,trifluoromethyl, C₁ -C₆ straight or branched alkyl or alkenyl, C₁ -C₄alkoxy or C₁ -C₄ alkenyloxy, phenoxy, benzyloxy, and amino;

X is oxygen, sulfur, methylene (CH₂) , or H₂ ;

Y is oxygen or NR₂, where R₂ is hydrogen or C₁ -C₆ alkyl; and

Z is a C₂ -C₆ straight or branched chain alkyl or alkenyl, wherein thealkyl chain is substituted in one or more positions with Ar₁ as definedabove, C₃ -C₈ cycloalkyl, cycloalkyl connected by a C₁ -C₆ straight orunbranched alkyl or alkenyl chain, or Ar₂ where Ar₂ is selected from thegroup consisting of 2-indolyl, 3-indolyl, 2-furyl, 3-furyl, 2-thiazolyl, 2-thienyl, 3-thienyl, 2-, 3-, or 4-pyridyl, or phenyl, havingone to three substituents which are independently selected from thegroup consisting of hydrogen, halo, hydroxyl, nitro, trifluoromethyl, C₁-C₆ straight or branched alkyl or alkenyl, C₁ -C₄ alkoxy or C₁ -C₄alkenyloxy, phenoxy, benzyloxy, and amino;

Z may also be the fragment: ##STR6## where R₃ is selected from the groupconsisting of straight or branched alkyl C₁ -C₈ optionally substitutedwith C₃ -C₈ cycloalkyl, or Ar₁ as defined above, and unsubstituted Ar₁ ;

X₂ is O or NR₅, where R₅ is selected from the group consisting ofhydrogen, C₁ -C₆ straight or branched alkyl and alkenyl;

R₄ is selected from the group consisting of phenyl, benzyl, C₁ -C₅straight or branched alkyl or alkenyl, and C₁ -C₅ straight or branchedalkyl or alkenyl substituted with phenyl; or pharmaceutically acceptablesalts or hydrates thereof.

Preferred compounds have the following formula: ##STR7## where R₁ is aC₁ -C₉ straight or branched chain alkyl or alkenyl group optionallysubstituted with C₃ -C₈ cycloalkyl, C₃ or C₅ cycloalkyl, C₅ -C₇cycloalkenyl, or Ar₁, where said alkyl, alkenyl, cycloalkyl orcycloalkenyl groups may be optionally substituted with C₁ -C₄ alkyl, C₁-C₄ alkenyl, or hydroxy, and where Ar₁ is selected from the groupconsisting of 1-napthyl, 2-napthyl, 2-indolyl, 3-indolyl, 2-furyl,3-furyl, 2- thiazolyl, 2-thienyl, 3-thienyl, 2-, 3-, or 4-pyridyl, orphenyl, having one to three substituents which are independentlyselected from the group consisting of hydrogen, halo, hydroxyl, nitro,trifluoromethyl, C₁ -C₆ straight or branched alkyl or alkenyl, C₁ -C₄alkoxy or C₁ -C₄ alkenyloxy, phenoxy, benzyloxy, and amino;

Z is a C₂ -C₆ straight or branched chain alkyl or alkenyl, wherein thealkyl chain is substituted in one or more positions with Ar₁ as definedabove, C3-C8 cycloalkyl, cycloalkyl connected by a C₁ -C₆ straight orunbranched alkyl or alkenyl chain, or Ar₂ where Ar₂ is selected from thegroup consisting of 2-indolyl, 3-indolyl, 2-furyl, 3-furyl, 2-thiazolyl, 2-thienyl, 3-thienyl, 2-, 3-, or 4-pyridyl, or phenyl, havingone to three substituents which are independently selected from thegroup consisting of hydrogen, halo, hydroxyl, nitro, trifluoromethyl, C₁-C₆ straight or branched alkyl or alkenyl, C₁ -C₄ alkoxy or C₁ -C₄alkenyloxy, phenoxy, benzyloxy, and amino; or pharmaceuticallyacceptable salts or hydrates thereof.

Preferred neurotrophic N-glyoxyl prolyl ester compounds have theformula: ##STR8## where R₁ is a C₁ -C₅ straight or branched chain alkylor alkenyl group optionally substituted with C₃ to C₆ cycloalkyl, orAr₁, where Ar₁ is selected from the group consisting of 2-furyl,2-thienyl, or phenyl;

X is selected from the group consisting of oxygen and sulfur;

Y is oxygen; and

Z is a straight or branched chain alkyl or alkenyl, wherein the alkylchain is substituted in one or more positions with Ar₁ as defined above,C₃ -C₆ cycloalkyl, Ar₂ where Ar₂ is selected from the group consistingof 2-, 3-, or 4-pyridyl, or phenyl, having one to three substituentswhich are independently selected from the group consisting of hydrogenand C₁ -C₄ alkoxy.

The compounds of this invention exist as stereoisomeric forms, eitherenantiomers or diastereoisomers. The stereochemistry at position 1(Formula 1) is R or S, with S preferred. Included within the scope ofthe invention are the enantiomers, the racemic form, anddiastereoisomeric mixtures. Enantiomers as well as diastereoisomers canbe separated by methods known to those skilled in the art.

It is known that immunophilins such as FKBP preferentially recognizepeptide substrates containing Xaa-Pro-Yaa motifs, where Xaa and Yaa arelipophilic amino acid residues. Schreiber et al. 1990 J. Org. Chem. 55,4984-4986; Harrison and Stein, 1990 Biochemistry, 29, 3813-3816. Thusmodified prolyl peptidomimetic compounds bearing lipophilic substituentsshould bind with high affinity to the hydrophobic core of the FKBPactive site and inhibit its rotamase activity.

Preferred compounds of the present invention include R₁ groups which arenot stereochemically bulky in relation to the known shape and size ofthe hydrophobic core of the FKBP active site. Thus, very large and/orhighly substituted R₁ groups would bind with less affinity to the FKBPactive site.

Preferred compounds of the invention include:

3-phenyl-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,

3-phenyl-1-prop-2-(E)-enyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,

3-(3,4,5-trimethoxyphenyl)-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,

3-(3,4,5-trimethoxyphenyl)-1-prop-2-(E)-enyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidine carboxylate,

3-(4,5-methylenedioxyphenyl)-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,

3-(4,5-methylenedioxyphenyl)-1-prop-2-(E)-enyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,

3-cyclohexyl-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,

3-cyclohexyl-1-prop-2-(E)-enyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,

(1R)-1,3-diphenyl-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,

3-phenyl-1-propyl (2S)-1-(1,2-dioxo-2-2-furanyl!)ethyl-2-pyrrolidinecarboxylate,

3-phenyl-1-propyl (2S)-1-(1,2-dioxo-2-2-thienyl!)ethyl-2-pyrrolidinecarboxylate,

3-phenyl-1-propyl (2S)-1-(1,2-dioxo-2-2-thiazolyl!)ethyl-2-pyrrolidinecarboxylate,

3-phenyl-1-propyl(2S)-1-(1,2-dioxo-2-phenyl)ethyl-2-pyrrolidinecarboxylate,

3-(2,5-dimethoxyphenyl)-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,

3-(2,5-dimethoxyphenyl)-1-prop-2-(E)-enyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,

2-(3,4,5-trimethoxyphenyl)-1-ethyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,

3-(3-Pyridyl)-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,

3-(2-Pyridyl)-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,

3-(4-Pyridyl)-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,

3-phenyl-1-propyl(2S)-1-(2-cyclohexyl-1,2-dioxoethyl)-2-pyrrolidinecarboxylate,

3-phenyl-1-propyl(2S)-1-(2-tert-butyl-1,2-dioxoethyl)-2-pyrrolidinecarboxylate,

3-phenyl-1-propyl(2S)-1-(2-cyclohexylethyl-1,2-dioxoethyl)-2-pyrrolidinecarboxylate,

3-(3-pyridyl)-1-propyl(2S)-1-(2-cyclohexylethyl-1,2-dioxoethyl)-2-pyrrolidinecarboxylate,

3-(3-pyridyl)-1-propyl(2S)-1-(2-tert-butyl-1,2-dioxoethyl)-2-pyrrolidinecarboxylate,

3,3-diphenyl-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,

3-(3-pyridyl)-1-propyl(2S)-1-(2-cyclohexyl-1,2-dioxoethyl)-2-pyrrolidinecarboxylate,

3-(3-Pyridyl)-1-propyl (2S)-N-( 2-thienyl!glyoxyl)pyrrolidinecarboxylate,

3,3-Diphenyl-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxobutyl)-2-pyrrolidinecarboxylate,

3,3-Diphenyl-1-propyl (2S)-1-cyclohexylglyoxyl-2-pyrrolidinecarboxylate,

3,3-Diphenyl-1-propyl(2S)-1-(2-thienyl)glyoxyl-2-pyrrolidinecarboxylate.

Particularly preferred neurotrophic N-glyoxyl prolyl ester compounds areselected from the group consisting of:

3-(2,5-dimethoxyphenyl)-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,

3-(2,5-dimethoxyphenyl)-1-prop-2-(E)-enyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,

2-(3,4,5-trimethoxyphenyl)-1-ethyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,

3-(3-Pyridyl)-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,

3-(2-Pyridyl)-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,

3-(4-Pyridyl)-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,

3-phenyl-1-propyl(2S)-1-(2-tert-butyl-1,2-dioxoethyl)-2-pyrrolidinecarboxylate,

3-phenyl-1-propyl(2S)-1-(2-cyclohexylethyl-1,2-dioxoethyl)-2-pyrrolidinecarboxylate,

3-(3-pyridyl)-1-propyl(2S)-1-(2-cyclohexylethyl-1,2-dioxoethyl)-2-pyrrolidinecarboxylate,

3-(3-pyridyl)-1-propyl(2S)-1-(2-tert-butyl-1,2-dioxoethyl)-2-pyrrolidinecarboxylate,

3,3-diphenyl-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,

3-(3-pyridyl)-1-propyl(2S)-1-(2-cyclohexyl-1,2-dioxoethyl)-2-pyrrolidinecarboxylate,

3-(3-Pyridyl)-1-propyl (2S)-N-( 2-thienyl!glyoxyl)pyrrolidinecarboxylate,

3,3-Diphenyl-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxobutyl)-2-pyrrolidinecarboxylate,

3,3-Diphenyl-1-propyl (2S)-1-cyclohexylglyoxyl-2-pyrrolidinecarboxylate,and

3,3-Diphenyl-1-propyl(2S)-1-(2-thienyl)glyoxyl-2-pyrrolidinecarboxylate.

The compounds of the present invention can be used in the form of saltsderived from inorganic or organic acids and bases. Included among suchacid salts are the following: acetate, adipate, alginate, aspartate,benzoate, benzenesulfonate, bisulfate butyrate, citrate, camphorate,camphorsulfonate, cyclopentanepropionate, digluconate, dodecylsulfate,ethanesulfonate, fumarate, glucoheptanoate, glycerophosphate,hemissulfate heptanoate, hexanoate, hydrochloride, hydrobromide,hydroiodide, 2-hydroxyethanesulfonate, lactate, maleate,methanesulfonate, 2-naphthalensulfonate, nicotinate, oxalate, pamoate,pectinate, propionate, succinate, tartrate, thiocyanate, tosylate andundecanoate. Base salts include ammonium salts, alkali metal salts suchas sodium and potassium salts, alkaline earth metal salts such ascalcium and magnesium salts, salt with organic bases such asdicyclohexylamine salts, N-methyl-D-glucamine, and salts with aminoacids such as arginine, lysine, and so forth. Also, the basicnitrogen-containing groups can be quarternized with such agents as loweralkyl halides, such as methyl, ethyl, propyl, and butyl chloride,bromides and iodides; dialkyl sulfates like dimethyl, diethyl, dibutyland diamyl sulfates, long chain halides such as decyl, lauryl, myristyland stearyl chlorides, bromides and iodides, aralkyl halides like benzyland phenethyl bromides and others. Water or oil-soluble or dispersibleproducts are thereby obtained.

The neurotrophic compounds of this invention can be periodicallyadministered to a patient undergoing treatment for neurologicaldisorders or for other reasons in which it is desirable to stimulateneuronal regeneration and growth, such as in various peripheralneuropathic and neurological disorders relating to neurodegeneration.The compounds of this invention can also be administered to mammalsother than humans for treatment of various mammalian neurologicaldisorders.

The novel compounds of the present invention are potent inhibitors ofrotamase activity and possess an excellent degree of neurotrophicactivity. This activity is useful in the stimulation of damaged neurons,the promotion of neuronal regeneration, the prevention ofneurodegeneration, and in the treatment of several neurologicaldisorders known to be associated with neuronal degeneration andperipheral neuropathies. The neurological disorders that may be treatedinclude but are not limited to: trigeminal neuralgia, glossopharyngealneuralgia, Bell's Palsy, myasthenia gravis, muscular dystrophy,amyotrophic lateral sclerosis, progressive muscular atrophy, progressivebulbar inherited muscular atrophy, herniated, ruptured or prolapsedinvertabrae disk syndromes, cervical spondylosis, plexus disorders,thoracic outlet destruction syndromes, peripheral neuropathic such asthose caused by lead, dapsone, ticks, prophyria, or Gullain-Barresyndrome, Alzheimer's disease, and Parkinson's disease.

For these purposes the compounds of the present invention may beadministered orally, parenterally, by inhalation spray, topically,rectally, nasally, buccally, vaginally or via an implanted reservoir indosage formulations containing conventional non-toxicpharmaceutically-acceptable carriers, adjuvants and vehicles. The termparenteral as used herein includes subcutaneous, intravenous,intramuscular, intraperitoneally, intrathecally, intraventricularly,intrasternal and intracranial injection or infusion techniques.

To be effective therapeutically as central nervous system targets theimmunophilin-drug complex should readily penetrate the blood-brainbarrier when peripherally administered. Compounds of this inventionwhich cannot penetrate the blood-brain barrier can be effectivelyadministered by an intraventricular route.

The pharmaceutical compositions may be in the form of a sterileinjectable preparation, for example as a sterile injectable aqueous oroleaginous suspension. This suspension may be formulated according totechniques know in the art using suitable dispersing or wetting agentsand suspending agents. The sterile injectable preparation may also be asterile injectable solution or suspension in a non-toxicparenterally-acceptable diluent or solvent, for example as a solution in1,3-butanediol. Among the acceptable vehicles and solvents that may beemployed are water, Ringer's solution and isotonic sodium chloridesolution. In addition, sterile, fixed oils are conventionally employedas a solvent or suspending medium. For this purpose any bland fixed oilmay be employed including synthetic mono- or diglycerides. Fatty acidssuch as oleic acid and its glyceride derivatives find use in thepreparation of injectables, olive oil or castor oil, especially in theirpolyoxyethylated versions. These oil solutions or suspensions may alsocontain a long-chain alcohol diluent or dispersant.

The compounds may be administered orally in the form of capsules ortablets, for example, or as an aqueous suspension or solution. In thecase of tablets for oral use, carriers which are commonly used includelactose and corn starch. Lubricating agents, such as magnesium stearate,are also typically added. For oral administration in a capsule form,useful diluents include lactose and dried corn starch. When aqueoussuspensions are required for oral use, the active ingredient is combinedwith emulsifying and suspending agents. If desired, certain sweeteningand/or flavoring and/or coloring agents may be added.

The compounds of this invention may also be administered in the form ofsuppositories for rectal administration of the drug. These compositionscan be prepared by mixing the drug with a suitable nonirritatingexcipient which is solid at room temperature but liquid at rectaltemperature and therefore will melt in the rectum to release the drug.Such materials include cocoa butter, beeswax and polyethylene glycols.

The compounds of this invention may also be administered optically,especially when the conditions addressed for treatment involve areas ororgans readily accessible by topical application, including neurologicaldisorders of the eye, the skin, or the lower intestinal tract. Suitabletopical formulations are readily prepared for each of these areas.

For ophthalmic use, the compounds can be formulated as micronizedsuspensions in isotonic, pH adjusted sterile saline, or, preferably, assolutions is isotonic, pH adjusted sterile saline, either with orwithout a preservative such as benzylalkonium chloride. Alternativelyfor the ophthalmic uses the compounds may be formulated in an ointmentsuch as petrolatum.

For application topically to the skin, the compounds can be formulatedin a suitable ointment containing the compound suspended or dissolvedin, for example, a mixture with one or more of the following: mineraloil, liquid petrolatum, white petrolatum, propylene glycol,polyoxyethylene polyoxypropylene compound, emulsifying wax and water.Alternatively, the compounds can be formulated in a suitable lotion orcream containing the active compound suspended or dissolved in, forexample, a mixture of one or more of the following: mineral oil,sorbitan monostearate, polysorbate 60, cetyl esters wax, cetearylalcohol, 2-octyldodecanol, benzyl alcohol and water.

Topical application for the lower intestinal tract an be effected in arectal suppository formulation (see above) or in a suitable enemaformulation.

Dosage levels on the order of about 0.1 mg to about 10,000 mg. of theactive ingredient compound are useful in the treatment of the aboveconditions, with preferred levels of about 0.1 mg to about 1,000 mg. Theamount of active ingredient that may be combined with the carriermaterials to produce a single dosage form will vary depending upon thehost treated and the particular mode of administration.

It is understood, however, that a specific dose level for any particularpatient will depend upon a variety of factors including the activity ofthe specific compound employed, the age, body weight, general health,sex, diet, time of administration, rate of excretion, drug combination,and the severity of the particular disease being treated and form ofadministration.

The compounds can be administered with other neurotrophic agents such asneurotrophic growth factor (NGF), glial derived growth factor, brainderived growth factor, ciliary neurotrophic factor, and neurotropin-3.The dosage level of other neurotrophic drugs will depend upon thefactors previously stated and the neurotrophic effectiveness of the drugcombination.

K_(i) Test Procedure

Inhibition of the peptidyl-prolyl isomerase (rotamase) activity of theinventive compounds can be evaluated by known methods described in theliterature (Harding, M. W. et al. Nature 341: 758-760 (1989); Holt etal. J. Am. Chem. Soc. 115: 9923-9938). These values are obtained asapparent K_(i) 's and are presented in Table I. The cis-transisomerization of an alanine-proline bond in a model substrate,N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide, is monitoredspectrophotometrically in a chymotrypsin-coupled assay, which releasespara-nitroanilide from the trans form of the substrate. The inhibitionof this reaction caused by the addition of different concentrations ofinhibitor is determined, and the data is analyzed as a change infirst-order rate constant as a function of inhibitor concentration toyield the apparent K_(i) values.

In a plastic cuvette are added 950 mL of ice cold assay buffer (25 mMHEPES, pH 7.8, 100 mM NaCl), 10 mL of FKBP (2.5 mM in 10 mM Tris-Cl pH7.5, 100 mM NaCl, 1 mM dithiothreitol), 25 mL of chymotrypsin (50 mg/mlin 1 mM HCl ) and 10 mL of test compound at various concentrations indimethyl sulfoxide. The reaction is initiated by the addition of 5 mL ofsubstrate (succinyl-Ala-Phe-Pro-Phe-para-nitroanilide, 5 mg/mL in 2.35mM LiCl in trifluoroethanol).

The absorbance at 390 nm versus time is monitored for 90 sec using aspectrophotometer and the rate constants are determined from theabsorbance versus time data files.

The data for these experiments is presented in Table I.

                  TABLE I    ______________________________________     ##STR9##    No.  R            R'                  K.sub.i    ______________________________________     1   1,1-dimethylpropyl                      3-phenylpropyl       42     2      "         3-phenyl-prop-2-(E)-enyl                                           125     3      "         3-(3,4,5-tri-methoxyphenyl)propyl                                           200     4      "         3-(3,4,5-trimethoxy-phenyl)-prop-2-                                           65                      (E)-enyl     5      "         3-(4,5-methylenedioxy)phenylpropyl                                           170     6      "         3-(4,5-methylenedioxy)phenylprop-2-                                           160                      (E)-enyl     7      "         3-cyclohexylpropyl   200     8      "         3-cyclohexylprop-2-(E)-enyl                                           600     9      "         (1R)-1,3-diphenyl-1-propyl                                           52    10   2-furanyl    3-phenylpropyl      4000    11   2-thienyl       "                 92    12   2-thiazolyl     "                 100    13   phenyl          "                1970    14   1,1-dimethylpropyl                      3-(2,5-dimethoxy)phenylpropyl                                           250    15      "         3-(2,5-dimethoxy)phenylprop-2-                                           450                      (E)-enyl    16      "         2-(3,4,5-trimethoxyphenyl)ethyl                                           120    17      "         3-(3-pyridyl)propyl   5    18                3-(2-pyridyl)propyl  195    19   1,1-dimethylpropyl                      3-(4-pyridyl)propyl  23    20   cyclohexyl   3-phenylpropyl       82    21   tert-butyl      "                 95    22   cyclohexylethyl                         "                1025    23   cyclohexylethyl                      3-(3-pyridyl)propyl 1400    24   tert-butyl   3-(3-pyridyl)propyl   3    25   1,1-dimethylpropyl                      3,3-diphenylpropyl    5    26   cyclohexyl   3-(3-pyridyl)propyl   9    27   2-thienyl    3-(3-pyridyl)propyl 1000    28   tert-butyl   3,3-diphenylpropyl    5    29   cyclohexyl      "                 20    30   2-thienyl       "                 150    ______________________________________

In mammalian cells, FKBP-12 complexes with the inositol triphosphatereceptor (IP₃ R) and the ryanodine receptor (RyR). It is believed thatthe neurotrophic compounds of this invention disassociates FKBP-12 fromthese complexes causing the calcium channel to become "leaky" (Cameronet al., 1995). Calcium fluxes are involved in neurite extensions so thatthe IP₃ R receptor and the ryanodine receptor might be involved in theneurotrophic effects of drugs. Since the drugs bind to the same site asFKBP-12 as the IP₃ R receptor, one could assume that the drugs displacethe channels from FKBP-12.

Chick Dorsal Root Ganglion Cultures and Neurite Outgrowth

Dorsal root ganglia were dissected from chick embryos of ten daygestation. Whole ganglion explants were cultured on thin layerMatrigel-coated 12 well plates with Liebovitz L15 plus high glucosemedia supplemented with 2 mM glutamine and 10% fetal calf serum, andalso containing 10 μM cytosine β-D arabinofuranoside (Ara C) at 37° C.in an environment containing 5% CO₂. Twenty-four hours later, the DRGswere treated with various concentrations of nerve growth factor,immunophilin ligands or combinations of NFG plus drugs. Forty-eighthours after drug treatment, the ganglia were visualized under phasecontrast or Hoffman Modulation contrast with a Zeiss Axiovert invertedmicroscope. Photomicrographs of the explants were made, and neuriteoutgrowth was quantitated. Neurites longer than the DRG diameter werecounted as positive, with total number of neurites quantitated per eachexperimental condition. Three to four DRGs are cultured per well, andeach treatment was performed in duplicate.

The data for these experiments are presented in Table II. Representativephotomicrographs for Example 17 are shown in FIG. 1; a dose responsecurve for this Example is given in FIG. 2.

                  TABLE II    ______________________________________    Neurite Outgrowth in Chick DRG    Example No. ED.sub.50, neurite outgrowth, nM    ______________________________________    1           53    2           105    3           149    4           190    5           10    6           75    10          0.46    11          0.015    14          2    15          0.8    16          0.015    17          0.05    18          30    19          6    20          0.13    21          0.025    22          0.66    23          1100    24          0.014    25          0.50    26          2    27          500    28          0.50    29          10    30          100    ______________________________________

Sciatic Nerve Axotomy

Six-week old male Sprague-Dawley rats were anesthetized, and the sciaticnerve exposed and crushed, at the level of the hip, by forceps. Testcompounds or vehicle were administered subcutaneously just prior to thelesion and daily for the following 18 days. Sections of the sciaticnerve were stained with Holmes silver stain to quantify the number ofaxons, and Luxol fast blue to quantify the level of myelination.Eighteen days after lesion, there was a significant decrease in thenumber of axons (50% decrease as compared to non-lesioned control) anddegree of myelination (90% decrease as compared to non-lesioned control)in animal treated with vehicle.

Administration of Example 1 (30 mg/kg s.c.), just prior to the lesionand daily for 18 days following the lesion, resulted in significantregeneration of both axon number (5% decrease as compared tonon-lesioned control) and the degree of myelination (50% decrease ascompared to control) as compared to vehicle treated animals. Thesignificant efficacy of Example 1 is consistent with its potent activityin inhibiting rotamase activity and stimulating neurite outgrowth inchick DRGs. These results are shown in FIG. 3. "Sham" denotes controlanimals that received vehicle but were not lesioned; "Vehicle" denotesanimals that were lesioned and received only vehicle (i.e., no drug).Example 1 showed a striking similarity to the sham treated animals,demonstrating the powerful neuroregenerative effects of these compoundsin vivo. Inactive is a compound that is inactive as an FKBP12 inhibitor.Animals treated with this compound resembled the vehicle-treatedlesioned animals, consistent with the neuroregenerative results observedwith Example 1 being directly caused by its inhibition of FKBP12.Quantitation for these data are shown in Table III.

                  TABLE III    ______________________________________                  Axon Number    Treatment     (% Control)                             Myelin Level    ______________________________________    Sham          100        100    Lesion:    +Vehicle (s.c.)                  50         10    +Example 1    100        50    (30 mg/    kg s.c.)    +Inactive     25         25    (30 mg/kg s.c.)    ______________________________________

MPTP Model of Parkinson's Disease in Mice

MPTP lesioning of dopaminergic neurons in mice was used as an animalmodel of Parkinson's Disease. Four week old male CD1 white mice weredosed i.p. with 30 mg/kg of MPTP for 5 days. Example 17(10-40 mg/kg), orvehicle, were administered s.c. along with the MPTP for 5 days, as wellas for an additional 5 days following cessation of MPTP treatment. At 18days following MPTP treatment, the animals were sacrificed and thestriata were dissected and homogenized. Binding of 3H!CFT, a radioligandfor the dopamine transporter, to the stiatal membranes was done toquantitate the level of the dopamine transporter (DAT) following lesionand drug treatment. Immunostaining was performed on saggital and coronalbrain sections using anti-tyrosine hydoxylase Ig to quantitate survivaland recovery of dopaminergic neurons. In animals treated with MPTP andvehicle, a substantial loss of functional dopaminergic terminals wasobserved as compared to non-lesioned animals. Lesioned animals receivingExample 17 showed a nearly quantitative recovery of TH-staineddopaminergic neurons.

FIGS. 4 and 5 show the quantitation in DAT levels, whereas FIGS. 6-8 arephotomicrographs showing the regenerative effects of Example 17 in thismodel. FIG. 4 demonstrates the significant recovery in functionaldopaminergic terminals, as assayed by 3H!-CFT binding, relative toanimals receiving MPTP but not the Guilford compounds. FIG. 5 gives thisdata in bar graph form. It is shown that animals receiving 40 mg/kg ofExample 17 in addition to MPTP manifested a greater than 90% recovery of3H!-CFT binding. As shown in FIGS. 6-8, immunostaining for tyrosinehydroxylase (a marker of viable dopaminergic neurons) in the striatum,the nigra, and the medial forebrain bundle, shows a clear and markedrecovery of functional neurons in animals that received Example 17, ascompared to animals that received lesioning agent but no drug(MPTP/Vehicle).

The following examples are illustrative of preferred embodiments of theinvention and are not to be construed as limiting the invention thereto.All preferred embodiments of the invention and are not to be construedas limiting the invention thereto. All polymer molecular weights aremean average molecular weights. All percentages are based on the percentby weight of the final delivery system or formulation prepared unlessotherwise indicated and all totals equal 100% by weight.

EXAMPLES

The inventive compounds may be prepared by a variety of syntheticsequences that utilize established chemical transformations. The generalpathway to the present compounds is described in Scheme 1.N-glyoxylproline derivatives may be prepared by reacting L-prolinemethyl ester with methyl oxalyl chloride as shown in Scheme I. Theresulting oxamates may be reacted with a variety of carbon nucleophilesto obtain intermediates compounds. These intermediates are then reactedwith a variety of alcohols, amides, or protected amino acid residues toobtain the propyl esters and amides of the invention. ##STR10##

Example 1

Synthesis of 3-phenyl-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate (Example1).

Synthesis of methyl(2S)-1-(1,2-dioxo-2-methoxyethyl)-2-pyrrolidinecarboxylate.

A solution of L-proline methyl ester hydrochloride (3.08 g; 18.60 mmol)in dry methylene chloride was cooled to 0° C. and treated withtriethylamine (3.92 g; 38.74 mmol; 2.1 eq). After stirring the formedslurry under a nitrogen atmosphere for 15 min, a solution of methyloxalyl chloride (3.20 g; 26.12 mmol) in methylene chloride (45 mL) wasadded dropwise. The resulting mixture was stirred at 0° C. for 1.5 hr.After filtering to remove solids, the organic phase was washed withwater, dried over MgSO₄ and concentrated. The crude residue was purifiedon a silica gel column, eluting with 50% ethyl acetate in hexane, toobtain 3.52 g (88%) of the product as a reddish oil. Mixture ofcis-trans amide rotamers; data for trans rotamer given. ¹ H NMR (CDCl₃): d 1.93 (dm, 2H); 2.17 (m, 2H) 3.62 (m, 2H); 3.71 (s, 3H); 3.79, 3.84(s, 3H total); 4.86 (dd, 1H, J=8.4, 3.3).

Synthesis of methyl(2S)-1-(1,2-dioxo-3,3-dimethylpentyl)-2-pyrrolidinecarboxylate.

A solution of methyl(2S)-1-(1,2-dioxo-2-methoxyethyl)-2-pyrrolidinecarboxylate (2.35 g;10.90 mmol) in 30 mL of tetrahydrofuran (THF) was cooled to -78° C. andtreated with 14.2 mL of a 1.0M solution of 1,1-dimethylpropylmagnesiumchloride in THF. After stirring the resulting homogeneous mixture at-78° C. for three hours, the mixture was poured into saturated ammoniumchloride (100 mL) and extracted into ethyl acetate. The organic phasewas washed with water, dried, and concentrated, and the crude materialobtained upon removal of the solvent was purified on a silica gelcolumn, eluting with 25% ethyl acetate in hexane, to obtain 2.10 g (75%)of the oxamate as a colorless oil. ¹ H NMR (CDCl₃) : d 0.88 (t, 3H) ;1.22, 1.26 (s, 3H each); 1.75 (dm, 2H); 1.87-2.10 (m, 3H); 2.23 (m, 1H);3.54 (m, 2H); 3.76 (s, 3H); 4.52 (dm, 1H, J=8.4, 3.4).

Synthesis of(2S)-1-(1,2-dioxo-3,3-dimethylpentyl)-2-pyrrolidinecarboxylic acid.

A mixture of methyl(2S)-1-(1,2-dioxo-3,3-dimethylpentyl)-2-pyrrolidinecarboxylate (2.10 g;8.23 mmol), 1N LiOH (15 mL), and methanol (50 mL) was stirred at 0° C.for 30 min and at room temperature overnight. The mixture was acidifiedto pH 1 with 1N HCl, diluted with water, and extracted into 100 mL ofmethylene chloride. The organic extract was washed with brine andconcentrated to deliver 1.73 g (87%) of snow-white solid which did notrequire further purification. ¹ H NMR (CDCl₃) : d 0.87 (t, 3H) ; 1.22,1.25 (s, 3H each); 1.77 (dm, 2H); 2.02 (m, 2H); 2.17 (m, 1H); 2.25 (m,1H); 3.53 (dd, 2H, J=10.4, 7.3); 4.55 (dd, 1H, J=8.6, 4.1).

Synthesis of 3-phenyl-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate (Example1). A mixture of(2S)-1-(1,2-dioxo-3,3-dimethylpentyl)-2-pyrrolidine-carboxylic acid (600mg; 2.49 mmol), 3-phenyl-1-propanol (508 mg; 3.73 mmol),dicyclohexylcarbodiimide (822 mg; 3.98 mmol), camphorsulphonic acid (190mg; 0.8 mmol) and 4-dimethylaminopyridine (100 mg; 0.8 mmol) inmethylene chloride (20 mL) was stirred overnight under a nitrogenatmosphere. The reaction mixture was filtered through Celite to removesolids and concentrated in vacuo, and the crude material was purified ona flash column (25% ethyl acetate in hexane) to obtain 720 mg (80%) ofExample 1 as a colorless oil. ¹ H NMR (CDCl₃): d 0.84 (t, 3H); 1.19 (s,3H); 1.23 (s, 3H); 1.70 (dm, 2H); 1.98 (m, 5H); 2.22 (m, 1H); 2.64 (m,2H); 3.47 (m, 2H); 4.14 (m, 2H); 4.51 (d, 1H); 7.16 (m, 3H); 7.26 (m,2H).

The method of Example 1 was utilized to prepare the followingillustrative examples:

Example 2

3-phenyl-1-prop-2-(E)-enyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate, 80%, ¹ HNMR (360 Mhz, CDCl₃) : d 0.86 (t, 3H) ; 1.21 (s, 3H); 1.25 (s, 3H);1.54-2.10 (m, 5H); 2.10-2.37 (m, 1H); 3.52-3.55 (m, 2H); 4.56 (dd, 1H,J=3.8, 8.9); 4.78-4.83 (m, 2H); 6.27 (m, 1H); 6.67 (dd, 1H, J=15.9);7.13-7.50 (m, 5H).

Example 3

3-(3,4,5-trimethoxyphenyl)-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate, 61%, ¹ HNMR (CDCl₃) : d 0.84 (t, 3H) ; 1.15 (s, 3H); 1.24 (s, 3H); 1.71 (dm,2H); 1.98 (m, 5H); 2.24 (m, 1H); 2.63 (m, 2H); 3.51 (t, 2H); 3.79 (s,3H); 3.83 (s, 3H); 4.14 (m, 2H); 4.52 (m, 1H); 6.36 (s, 2H).

Example 4

3-(3,4,5-trimethoxyphenyl)-1-prop-2-(E)-enyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidine carboxylate, 66%, ¹H NMR (CDCl₃) : d 0.85 (t, 3H) ; 1.22 (s, 3H); 1.25 (s, 3H); 1.50-2.11(m, 5H); 2.11-2.40 (m, 1H); 3.55 (m, 2H); 3.85 (s, 3H); 3.88 (s, 6H);4.56 (dd, 1H); 4.81 (m, 2H); 6.22 (m, 1H); 6.58 (d, 1H, J=16); 6.63 (s,2H).

Example 5

3-(4,5-methylenedioxyphenyl)-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate, 82%, ¹ HNMR (360 MHz, CDCl₃) : d 0.86 (t,3H); 1.22 (s, 3H); 1.25 (s, 3H);1.60-2.10 (m, 5H); 3.36-3.79 (m, 2H); 4.53 (dd, 1H, J=3.8, 8.6);4.61-4.89 (m, 2H); 5.96 (s, 2H); 6.10 (m, 1H); 6.57 (dd, 1H, J=6.2,15.8); 6.75 (d, 1H, J=8.0); 6.83 (dd, 1H, J=1.3, 8.0); 6.93 (s, 1H)

Example 6

3-(4,5-methylenedioxyphenyl)-1-prop-2-(E)-enyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,82%, ¹ HNMR (360 MHz, CDCl₃) : d 0.86 (t, 3H); 1.22 (s, 3H); 1.25 (s, 3H);1.60-2.10 (m, 5H); 2.10-2.39 (m, 1H); 3.36-3.79 (m, 2H); 4.53 (dd, 1H,J=3.8, 8.6); 4.61-4.89 (m, 2H); 5.96 (s, 2H); 6.10 (m, 1H); 6.57 (dd,1H, J=6.2, 15.8); 6.75 (d, 1H, J=8.0); 6.83 (dd, 1H, J=1.3, 8.0); 6.93(s, 1H).

Example 8

3-cyclohexyl-1-prop-2-(E)-enyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate, 92%, ¹ HNMR (360 MHz, CDCl₃) : d 0.86 (t, 3H) ; 1.13-1.40 (m+2 singlets, 9Htotal); 1.50-1.87 (m, 8H); 1.87-2.44 (m, 6H); 3.34-3.82 (m, 2H);4.40-4.76 (m, 3H); 5.35-5.60 (m, 1H); 5.60-5.82 (dd, 1H, J=6.5, 16).

Example 9

(1R)-1,3-Diphenyl-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate, 90%, ¹ HNMR (360 MHz, CDCl₃) : d 0.85 (t, 3H) ; 1.20 (s, 3H); 1.23 (s, 3H);1.49-2.39 (m, 7H); 2.46-2.86 (m, 2H); 3.25-3.80 (m, 2H); 4.42-4.82 (m,1H); 5.82 (td, 1H, J=1.8, 6.7); 7.05-7.21 (m, 3H); 7.21-7.46 (m, 7H).

Example 10

3-phenyl-1-propyl (2S)-1-(1,2-dioxo-2-2-furanyl!)ethyl-2-pyrrolidinecarboxylate, 99%, ¹ H NMR (300 MHz, CDCl₃): d 1.66-2.41 (m, 6H) ; 2.72 (t, 2H, J=7.5); 3.75 (m, 2H); 4.21 (m, 2H);4.61 (m, 1H); 6.58 (m, 1H); 7.16-7.29 (m, 5H); 7.73 (m, 2H).

Example 11

3-phenyl-1-propyl (2S)-1-(1,2-dioxo-2-2-thienyl!)ethyl-2-pyrrolidinecarboxylate, 81%, ¹ H NMR (300 MHz, CDCl₃): d 1.88-2.41 (m, 6H) ; 2.72 (dm, 2H) 3.72 (m, 2H); 4.05 (m, 1H); 4.22(m, 1H); 4.64 (m, 1H); 7.13-7.29 (m, 6H); 7.75 (dm, 1H); 8.05 (m, 1H).

Example 13

3-phenyl-1-propyl(2S)-1-(1,2-dioxo-2-phenyl)ethyl-2-pyrrolidinecarboxylate, 99%, ¹ H NMR(300 MHz, CDCl₃) : d 1.97-2.32 (m, 6H) ; 2.74 (t, 2H, J=7.5); 3.57 (m,2H); 4.24 (m, 2H); 4.67 (m, 1H); 6.95-7.28 (m, 5H); 7.51-7.64 (m, 3H);8.03-8.09 (m, 2H).

Example 14

3-(2,5-dimethoxyphenyl)-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate, 99%, ¹ HNMR (300 MHz, CDCl₃) : d 0.87 (t, 3H); 1.22 (s, 3H); 1.26 (s, 3H); 1.69(m, 2H); 1.96 (m, 5H); 2.24 (m, 1H); 2.68 (m, 2H); 3.55 (m, 2H); 3.75(s, 3H); 3.77 (s, 3H); 4.17 (m, 2H); 4.53 (d, 1H); 6.72 (m, 3H).

Example 15

3-(2,5-dimethoxyphenyl)-1-prop-2-(E)-enyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate, 99%, ¹ HNMR (300 MHz, CDCl₃) : d 0.87 (t, 3H); 1.22 (s, 3H); 1.26 (s, 3H); 1.67(m, 2H); 1.78 (m, 1H); 2.07 (m, 2H); 2.26 (m, 1H); 3.52 (m, 2H); 3.78(s, 3H); 3.80 (s, 3H); 4.54 (m, 1H); 4.81 (m, 2H); 6.29 (dt, 1H,J=15.9); 6.98 (s, 1H)

Example 16

2-(3,4,5-trimethoxyphenyl)-1-ethyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate, 97%, ¹ HNMR (300 MHz, CDCl₃) : d 0.84 (t, 3H); 1.15 (s, 3H); 1.24 (s, 3H); 1.71(dm, 2H); 1.98 (m, 5H); 2.24 (m, 1H); 2.63 (m, 2H); 3.51 (t, 2H); 3.79(s, 3H); 3.83 (s, 3H); 4.14 (m, 2H); 4.52 (m, 1H); 6.36 (s, 2H).

Example 17

3-(3-Pyridyl)-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate, 80%, ¹ HNMR (CDCl₃, 300 MHz) : d 0.85 (t, 3H) ; 1.23, 1.26 (s, 3H each);1.63-1.89 (m, 2H); 1.90-2.30 (m, 4H); 2.30-2.50 (m, 1H); 2.72 (t, 2H);3.53 (m, 2H); 4.19 (m, 2H); 4.53 (m, 1H); 7.22 (m, 1H); 7.53 (dd, 1H);8.45.

Example 18

3-(2-Pyridyl)-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate, 88%, ¹ HNMR (CDCl₃, 300 MHz) : d 0.84 (t, 3H) ; 1.22, 1.27 (s, 3H each);1.68-2.32 (m, 8H); 2.88 (t, 2H, J=7.5); 3.52 (m, 2H); 4.20 (m, 2H); 4.51(m, 1H); 7.09-7.19 (m, 2H); 7.59 (m, 1H); 8.53 (d, 1H, J=4.9).

Example 19

3-(4-Pyridyl)-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate, 91%, ¹ HNMR (CDCl₃, 300 MHz) : d 6.92-6.80 (m, 4H) ; 6.28 (m, 1H); 5.25 (d, 1H,J=5.7); 4.12 (m, 1H); 4.08 (s, 3H); 3.79 (s, 3H); 3.30 (m, 2H); 2.33 (m,1H); 1.85-1.22 (m, 7H); 1.25 (s, 3H); 1.23 (s, 3H); 0.89 (t, 3H, J=7.5).

Example 20

3-phenyl-1-propyl(2S)-1-(2-cyclohexyl-1,2-dioxoethyl)-2-pyrrolidinecarboxylate, 91%, ¹ HNMR (CDCl₃, 300 MHz) : d 1.09-1.33 (m, 5H) ; 1.62-2.33 (m, 12H); 2.69)t, 2H, J=7.5); 3.15 (dm, 1H); 3.68 (m, 2H); 4.16 (m, 2H); 4.53, 4.84(d, 1H total); 7.19 (m, 3H); 7.29 (m, 2H).

Example 21

3-phenyl-1-propyl(2S)-1-(2-tert-butyl-1,2-dioxoethyl)-2-pyrrolidinecarboxylate, 92%, ¹ HNMR (CDCl₃, 300 MHz) : d 1.29 (s, 9H) ; 1.94-2.03 (m, 5H) 2.21 (m, 1H);2.69 (m, 2H); 3.50-3.52 (m, 2H); 4.16 (m, 2H); 4.53 (m, 1H); 7.19 (m,3H); 7.30 (m, 2H).

Example 22

3-phenyl-1-propyl(2S)-1-(2-cyclohexylethyl-1,2-dioxoethyl)-2-pyrrolidinecarboxylate, 97%,¹ H NMR (CDCl₃, 300 MHz) : d 0.88 (m, 2H) ; 1.16 (m, 4H); 1.43-1.51 (m,2H); 1.67 (m, 5H); 1.94-2.01 (m, 6H); 2.66-2.87 (m, 4H); 3.62-3.77 (m,2H); 4.15 (m, 2H); 4.86 (m, 1H); 7.17-7.32 (m, 5H).

Example 23

3-(3-pyridyl)-1-propyl(2S)-1-(2-cyclohexylethyl-1,2-dioxoethyl)-2-pyrrolidinecarboxylate, 70%,¹ H NMR (CDCl₃, 300 MHz) : d 0.87 (m, 2H) ; 1.16 (m, 4H); 1.49 (m, 2H);1.68 (m, 4H); 1.95-2.32 (m, 7H); 2.71 (m, 2H); 2.85 (m, 2H); 3.63-3.78(m, 2H); 4.19 (m, 2H); 5.30 (m, 1H); 7.23 (m, 1H); 7.53 (m, 1H); 8.46(m, 2H).

Example 24

3-(3-pyridyl)-1-propyl(2S)-1-(2-tert-butyl-1,2-dioxoethyl)-2-pyrrolidinecarboxylate, 83%, ¹ HNMR (CDCl₃, 300 MHz) : d 1.29 (s, 9H) ; 1.95-2.04 (m, 5H); 2.31 (m, 1H);2.72 (t, 2H, J=7.5); 3.52 (m, 2H); 4.18 (m, 2H) ; 4.52 (m, 1H) ;7.19-7.25 (m, 1H) ; 7.53 (m, 1H) ; 8.46 (m, 2H).

Example 25

3,3-diphenyl-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate, 99%, ¹ HNMR (CDCl₃, 300 MHz) : d 0.85 (t, 3H) ; 1.21, 1.26 (s, 3H each);1.68-2.04 (m, 5H); 2.31 (m, 1H); 2.40 (m, 2H); 3.51 (m, 2H); 4.08 (m,3H); 4.52 (m, 1H); 7.18-7.31 (m, 10H).

Example 26

3-(3-pyridyl)-1-propyl(2S)-1-(2-cyclohexyl-1,2-dioxoethyl)-2-pyrrolidinecarboxylate, 88%, ¹ HNMR (CDCl₃, 300 MHz) : d 1.24-1.28 (m, 5H) ; 1.88-2.35 (m, 11H); 2.72(t, 2H, J=7.5); 3.00-3.33 (dm, 1H); 3.69 (m, 2H); 4.19 (m, 2H); 4.55 (m,1H); 7.20-7.24 (m, 1H); 7.53 (m, 1H); 8.47 (m, 2H).

Example 27

3-(3-Pyridyl)-1-propyl (2S)-N-( 2-thienyl!glyoxyl)pyrrolidinecarboxylate, 49%, ¹ H NMR (CDCl₃, 300 MHz): d1.81-2.39 (m, 6H); 2.72 (dm, 2H); 3.73 (m, 2H); 4.21 (m, 2H); 4.95 (m,1H); 7.19 (m, 2H); 7.61 (m, 1H); 7.80 (d, 1H); 8.04 (d, 1H); 8.46 (m,2H).

Example 28

3,3-Diphenyl-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxobutyl)-2-pyrrolidinecarboxylate, 99%, ¹ HNMR (CDCl₃, 300 MHz) : d 1.27 (s, 9H) ; 1.96 (m, 2H) ; 2.44 (m, 4H);3.49 (m, 1H); 3.64 (m, 1H); 4.08 (m, 4H); 4.53 (dd, 1H) ; 7.24 (m, 10H).

Example 29

3,3-Diphenyl-1-propyl (2S)-1-cyclohexylglyoxyl-2-pyrrolidinecarboxylate, 91%, ¹ H NMR (CDCl₃, 300 MHz): d 1.32(m, 6H); 1.54-2.41 (m, 10H); 3.20 (dm, 1H); 3.69 (m, 2H); 4.12 (m, 4H);4.52 (d, 1H); 7.28 (m, 10H).

Example 30

3,3-Diphenyl-1-propyl (2S)-1-(2-thienyl)glyoxyl-2-pyrrolidinecarboxylate, 75%, ¹ H NMR (CDCl₃, 300 MHz): d 2.04(m, 3H); 2.26 (m, 2H); 2.48 (m, 1H); 3.70 (m, 2H); 3.82-4.18 (m, 3Htotal); 4.64 (m, 1H); 7.25 (m, 11H); 7.76 (dd, 1H); 8.03 (m, 1H).

The requisite substituted alcohols may be prepared by a number ofmethods known to those skilled in the art of organic synthesis. Asdescribed in Scheme II, alkyl or aryl aldehydes may be homologated tophenyl propanols by reaction with methyl(triphenylphosphoranylidene)acetate to provide a variety oftrans-cinnamates; these latter may be reduced to the saturated alcoholsby reaction with excess lithium aluminum hydride, or sequentially byreduction of the double bond by catalytic hydrogenation and reduction ofthe saturated ester by appropriate reducing agents. Alternatively, thetrans-cinnamates may be reduced to (E)-allylic alcohols by the use ofdiisobutylaluminum hydride. ##STR11##

Longer chain alcohols may be prepared by homologation of benzylic andhigher aldehydes. Alternatively, these aldehydes may be prepared byconversion of the corresponding phenylacetic and higher acids, andphenethyl and higher alcohols.

General procedure for the synthesis of acrylic esters, exemplified formethyl (3,3,5-trimethoxy)-trans-cinnamate:

A solution of 3,4,5-trimethoxybenzaldehyde (5.0 g; 25.48 mmol) andmethyl (triphenylphosphoranylidene)acetate (10.0 g; 29.91 mmol) intetrahydrofuran (250 mL) was refluxed overnight. After cooling, thereaction mixture was diluted with 200 mL of ethyl acetate and washedwith 2×200 mL of water, dried, and concentrated in vacuo. The cruderesidue was chromatographed on a silica gel column, eluting with 25%ethyl acetate in hexane, to obtain 5.63 g (88%) of the cinnamate as awhite crystalline solid, ¹ H NMR (300 Mhz; CDCl₃): d 3.78 (s, 3H); 3.85(s, 6H); 6.32 (d, 1H, J=16); 6.72 (s, 2H); 7.59 (d, 1H, J=16).

General procedure for the synthesis of saturated alcohols from acrylicesters. Exemplified for (3,4,5-trimethoxy) phenylpropanol.

A solution of methyl (3,3,5-trimethoxy)-trans-cinnamate (1.81 g; 7.17mmol) in tetrahydrofuran (30 mL) was added in a dropwise manner to asolution of lithium aluminum hydride (14 mmol) in THF (35 mL), withstirring and under an argon atmosphere. After the addition was complete,the mixture was heated to 75° C. for 4 hours. After cooling, it wasquenched by the careful addition of 15 mL of 2N NaOH followed by 50 mLof water. The resulting mixture was filtered through Celite to removesolids, and the filter cake was washed with ethyl acetate. The combinedorganic fractions were washed with water, dried, concentrated in vacuo,and purified on a silica gel column, eluting with ethyl acetate toobtain 0.86 g (53%) of the alcohol as a clear oil, ¹ H NMR (300 Mhz;CDCl₃): d 1.23 (br, 1H); 1.87 (m, 2H); 2.61 (t, 2H, J=7.1); 3.66 (t,2H); 3.80 (s, 3H); 3.83 (s, 6H); 6.40 (s, 2H).

General procedure for the synthesis of trans-allylic alcohols fromacrylic esters. Exemplified for (3,4,5-trimethoxy)phenylprop-2-(E)-enol.

A solution of methyl (3,3,5-trimethoxy)-trans-cinnamate (1.35 g; 5.35mmol) in toluene (25 mL) was cooled to -10° C. and treated with asolution of diisobutylaluminum hydride in toluene (11.25 mL of a 1.0Msolution; 11.25 mmol). The reaction mixture was stirred for 3 hrs at 0°C. and then quenched with 3 mL of methanol followed by 1N HCl until thepH was 1. The reaction mixture was extracted into ethyl acetate and theorganic phase was washed with water, dried and concentrated.Purification on a silica gel column eluting with 25% ethyl acetate inhexane furnished 0.96 g (80%) of a thick oil, ¹ H NMR (360 Mhz; CDCl₃):d 3.85 (s, 3H); 3.87 (s, 6H); 4.32 (d, 2H, J=5.6); 6.29 (dt, 1H, J=15.8,5.7), 6.54 (d, 1H, J=15.8); 6.61 (s, 2H).

The invention being thus described, it will be obvious that the same maybe varied in many ways. Such variations are not to be regarded as adeparture from the spirit and scope of the invention and all suchmodification are intended to be included within the scope of thefollowing claims.

What is claimed is:
 1. A non-immunosuppressive compound of the formula:##STR12## where R₁ is a C₁ -C₆ straight or branched chain alkyl oralkenyl group optionally substituted with C₃ to C₆ cycloalkyl, C₃ -C₆cycloalkyl, or Ar₁, where Ar₁ is selected from the group consisting of2-furyl, 2-thienyl, or phenyl;X is selected from the group consisting ofoxygen and sulfur; Y is oxygen; and Z is a C₂ -C₆ straight chain alkylor alkenyl, wherein the alkyl or alkenyl chain is substituted in one ormore positions with 2-furyl, 2-thienyl, C₃ -C₆ cycloalkyl, pyridyl, orphenyl, each optionally having one to three C₁ -C₄ alkoxy substituentsor pharmaceutically acceptable salt, hydrate, or mixture thereof.
 2. Thecompound of claim 1, where Z and R₁ are lipophilic groups.
 3. Anon-immunosuppresive compound selected from the group consistingof:3-(2,5-dimethoxyphenyl)-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,3-(2,5-dimethoxyphenyl)-1-prop-2-(E)-enyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,2-(3,4,5-trimethoxyphenyl)-1-ethyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,3-(3-Pyridyl)-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,3-(2-Pyridyl)-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,3-(4-Pyridyl)-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,3-phenyl-1-propyl(2S)-1-(2-tert-butyl-1,2-dioxoethyl)-2-pyrrolidinecarboxylate,3-phenyl-1-propyl(2S)-1-(2-cyclohexylethyl-1,2-dioxoethyl)-2-pyrrolidinecarboxylate,3-(3-pyridyl)-1-propyl(2S)-1-(2-cyclohexylethyl-1,2-dioxoethyl)-2-pyrrolidinecarboxylate,3-(3-pyridyl)-1-propyl(2S)-1-(2-tert-butyl-1,2-dioxoethyl)-2-pyrrolidinecarboxylate,3,3-diphenyl-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,3-(3-pyridyl)-1-propyl(2S)-1-(2-cyclohexyl-1,2-dioxoethyl)-2-pyrrolidinecarboxylate,3-(3-Pyridyl)-1-propyl (2S)-N-( 2-thienyl!glyoxyl)pyrrolidinecarboxylate, 3,3-Diphenyl-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxobutyl)-2-pyrrolidinecarboxylate,3,3-Diphenyl-1-propyl (2S)-1-cyclohexylglyoxyl-2-pyrrolidinecarboxylate,and 3,3-Diphenyl-1-propyl(2S)-1-(2-thienyl)glyoxyl-2-pyrrolidinecarboxylate and pharmaceuticallyacceptable salt, hydrate, and mixture thereof.
 4. A pharmaceuticalcomposition comprising a neurotrophically effective amount of a compoundof the formula: ##STR13## where R₁ is a C₁ -C₆ straight or branchedchain alkyl or alkenyl group optionally substituted with C₃ to C₆cycloalkyl, C₃ -C₆ cycloalkyl, or Ar₁, where Ar₁ is selected from thegroup consisting of 2-furyl, 2-thienyl, or phenyl;X is selected from thegroup consisting of oxygen and sulfur; Y is oxygen; and Z is a C₂ -C₆straight chain alkyl or alkenyl, wherein the alkyl or alkenyl chain issubstituted in one or more positions with 2-furyl, 2-thienyl, C₃ -C₆cycloalkyl, pyridyl, or phenyl, each optionally having one to three C₁-C₄ alkoxy substituents or pharmaceutically acceptable salt, hydrate, ormixture thereof,and a pharmaceutically acceptable carrier.
 5. A methodof stimulating growth of damaged nerves, which comprises:administeringto damaged nerves a compound of the formula: ##STR14## where R₁ is a C₁-C₆ straight or branched chain alkyl or alkenyl group optionallysubstituted with C₃ to C₆ cycloalkyl, C₃ -C₆ cycloalkyl, or Ar₁, whereAr₁ is selected from the group consisting of 2-furyl, 2-thienyl, orphenyl; X is selected from the group consisting of oxygen and sulfur; Yis oxygen; and Z is a C₂ -C₆ straight chain alkyl or alkenyl, whereinthe alkyl or alkenyl chain is substituted in one or more positions with2-furyl, 2-thienyl, C₃ -C₆ cycloalkyl, pyridyl, or phenyl, eachoptionally having one to three C₁ -C₄ alkoxy substituents orpharmaceutically acceptable salt, hydrate, or mixture thereof,insufficient amounts to stimulate the growth of said nerves.
 6. A methodof treating a neurological disorder selected from the group consistingof peripheral neuropathies, and neurological pathologies related toneurodegeneration in an animal which comprises administering atherapeutically effective amount of a compound of the formula: ##STR15##where R₁ is a C₁ -C₆ straight or branched chain alkyl or alkenyl groupoptionally substituted with C₃ to C₆ cycloalkyl, C₃ -C₆ cycloalkyl, orAr₁, where Ar₁ is selected from the group consisting of 2-furyl,2-thienyl, or phenyl;X is selected from the group consisting of oxygenand sulfur; Y is oxygen; and Z is a C₂ -C₆ straight chain alkyl oralkenyl, wherein the alkyl or alkenyl chain is substituted in one ormore positions with 2-furyl, 2-thienyl, C₃ -C₆ cycloalkyl, pyridyl, orphenyl, each optionally having one to three C₁ -C₄ alkoxy substituentsor pharmaceutically acceptable salt, hydrate, or mixture thereof,wheresaid compound has an affinity for FKBP-type immunophilins wherein theimmunophilin exhibits rotamase activity and the compound inhibits therotamase activity of the immunophilin.
 7. The method of claim 6, whereinthe FKBP-type immunophilin is FKBP-12.
 8. The method of claim 6, whereinthe neurological disorder is Alzheimer's disease.
 9. The method of claim6, wherein the neurological disorder is Parkinson's disease.
 10. Themethod of claim 6, wherein the neurological disorder is amyotrophiclateral sclerosis.
 11. The composition of claim 4 wherein the compoundis administered orally.
 12. The method of claim 5 wherein the compoundis administered orally.
 13. The method of claim 6 wherein the compoundis administered orally.
 14. A compound selected from the groupconsistingof:3-(3-Pyridyl)-1-propyl(2S)-1(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate;3-(2-Pyridyl)-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate;and, 3-(3-Pyridyl)-1-propyl(2S)-1-(2-cyclohexyl-1,2-dioxoethyl)-2-pyrrolidinecarboxylate or apharmaceutical salt or hydrate thereof. 15.3-(3-Pyridyl)-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,or a pharmaceutical salt, hydrate, or a mixture thereof.
 16. A method ofstimulating growth of damaged nerves, which comprises:administering todamaged nerves a compound in sufficient amounts to stimulate the growthof said nerves, wherein the compound is selected from the groupconsisting of:3-(2,5-dimethoxyphenyl)-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,3-(2,5-dimethoxyphenyl)-1-prop-2-(E)-enyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,2-(3,4,5-trimethoxyphenyl)-1-ethyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,3-(3-Pyridyl)-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,3-(2-Pyridyl)-1-propyl(2S)-1(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,3-(4-Pyridyl)-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,3-phenyl-1-propyl(2S)-1-(2-tert-butyl-1,2-dioxoethyl)-2-pyrrolidinecarboxylate,3-phenyl-1-propyl(2S)-1-(2-cyclohexylethyl-1,2-dioxoethyl)-2-pyrrolidinecarboxylate,3-(3-pyridyl)-1-propyl(2S)-1-(2-cyclohexylethyl-1,2-dioxoethyl)-2-pyrrolidinecarboxylate,3-(3-pyridyl)-1-propyl(2S)-1-(2-tert-butyl-1,2-dioxoethyl)-2-pyrrolidinecarboxylate,3,3-diphenyl-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,3-(3-pyridyl)-1-propyl(2S)-1-(2-cyclohexyl-1,2-dioxoethyl)-2-pyrrolidinecarboxylate,3-(3-Pyridyl)-1-propyl (2S)-N-( 2-thienyl!glyoxyl)pyrrolidinecarboxylate,3,3-Diphenyl-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxobutyl)-2-pyrrolidinecarboxylate,3,3-Diphenyl-1-propyl (2S)-1-cyclohexylglyoxyl-2-pyrrolidinecarboxylate,3,3-Diphenyl-1-propyl(2S)-1-(2-thienyl)glyoxyl-2-pyrrolidinecarboxylate, and apharmaceutically acceptable salt, hydrate, or a mixture thereof.
 17. Amethod of treating a neurological disorder selected from the groupconsisting of peripheral neuropathies, and neurological pathologiesrelated to neurodegeneration in an animal which comprises administeringa therapeutically effective amount of a compound selected from the groupconsisting of:3-(2,5-dimethoxyphenyl)-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,3-(2,5-dimethoxyphenyl)-1-prop-2-(E)-enyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,2-(3,4,5-trimethoxyphenyl)-1-ethyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,3-(3-Pyridyl)-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,3-(2-Pyridyl)-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,3-(4-Pyridyl)-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,3-phenyl-1-propyl(2S)-1-(2-tert-butyl-1,2-dioxoethyl)-2-pyrrolidinecarboxylate,3-phenyl-1-propyl(2S)-1-(2-cyclohexylethyl-1,2-dioxoethyl)-2-pyrrolidinecarboxylate,3-(3-pyridyl)-1-propyl(2S)-1-(2-cyclohexylethyl-1,2-dioxoethyl)-2-pyrrolidinecarboxylate,3-(3-pyridyl)-1-propyl(2S)-1-(2-tert-butyl-1,2-dioxoethyl)-2-pyrrolidinecarboxylate,3,3-diphenyl-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,3-(3-pyridyl)-1-propyl(2S)-1-(2-cyclohexyl-1,2-dioxoethyl)-2-pyrrolidinecarboxylate,3-(3-Pyridyl)-1-propyl (2S)-N-( 2-thienyl! glyoxyl)pyrrolidinecarboxylate,3,3-Diphenyl-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxobutyl)-2-pyrrolidinecarboxylate,3,3-Diphenyl-1-propyl (2S)-1-cyclohexylglyoxyl-2-pyrrolidinecarboxylate,3. 3-Diphenyl-1-propyl(2S)-1-(2-thienyl)glyoxyl-2-pyrrolidinecarboxylate, andapharmaceutically acceptable salt, hydrate, or a mixture thereof, wheresaid compound has an affinity for FKBP-type immunophilins wherein theimmunophilin exhibits rotamase activity and the compound inhibits therotamase activity of the immunophilin.
 18. A pharmaceutical compositioncomprising:i) a neurotrophically effective amount of a compound selectedfrom the group consisting of:3-(2,5-dimethoxyphenyl)-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,3-(2,5-dimethoxyphenyl)-1-prop-2-(E)-enyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,2-(3,4,5-trimethoxyphenyl)-1-ethyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,3-(3-Pyridyl)-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,3-(2-Pyridyl)-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,3-(4-Pyridyl)-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,3-phenyl-1-propyl(2S)-1-(2-tert-butyl-1,2-dioxoethyl)-2-pyrrolidinecarboxylate,3-phenyl-1-propyl(2S)-1-(2-cyclohexylethyl-1,2-dioxoethyl)-2-pyrrolidinecarboxylate,3-(3-pyridyl)-1-propyl(2S)-1-(2-cyclohexylethyl-1,2-dioxoethyl)-2-pyrrolidinecarboxylate,3-(3-pyridyl)-1-propyl(2S)-1-(2-tert-butyl-1,2-dioxoethyl)-2-pyrrolidinecarboxylate,3,3-diphenyl-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,3-(3-pyridyl)-1-propyl(2S)-1-(2-cyclohexyl-1,2-dioxoethyl)-2-pyrrolidinecarboxylate,3-(3-Pyridyl)-1-propyl (2S)-N-( 2-thienyl! glyoxyl)pyrrolidinecarboxylate,3,3-Diphenyl-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxobutyl)-2-pyrrolidinecarboxylate,3,3-Diphenyl-1-propyl (2S)-1-cyclohexylglyoxyl-2-pyrrolidinecarboxylate,3,3-Diphenyl-1-propyl(2S)-1-(2-thienyl)glyoxyl-2-pyrrolidinecarboxylate, and apharmaceutically acceptable salt, hydrate, or a mixture thereof; and ii)a pharmaceutically acceptable carrier.
 19. A method of treatingParkinson's disease in an animal which comprises administering atherapeutically effective amount of a compound selected from the groupconsisting of:3-(2,5-dimethoxyphenyl)-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,3-(2,5-dimethoxyphenyl)-1-prop-2-(E)-enyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,2-(3,4,5-trimethoxyphenyl)-1-ethyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,3-(3-Pyridyl)-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,3-(2-Pyridyl)-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,3-(4-Pyridyl)-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,3-phenyl-1-propyl(2S)-1-(2-tert-butyl-1,2-dioxoethyl)-2-pyrrolidinecarboxylate,3-phenyl-1-propyl(2S)-1-(2-cyclohexylethyl-1,2-dioxoethyl)-2-pyrrolidinecarboxylate,3-(3-pyridyl)-1-propyl(2S)-1-(2-cyclohexylethyl-1,2-dioxoethyl)-2-pyrrolidinecarboxylate,3-(3-pyridyl)-1-propyl(2S)-1-(2-tert-butyl-1,2-dioxoethyl)-2-pyrrolidinecarboxylate,3,3-diphenyl-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate,3-(3-pyridyl)-1-propyl(2S)-1-(2-cyclohexyl-1,2-dioxoethyl)-2-pyrrolidinecarboxylate,3-(3-Pyridyl)-1-propyl (2S)-N-( 2-thienyl! glyoxyl)pyrrolidinecarboxylate, 3.3-Diphenyl-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxobutyl)-2-pyrrolidinecarboxylate,3,3-Diphenyl-1-propyl(2S)-1-cyclohexylglyoxyl-2-pyrrolidinecarboxylate, 3,3-Diphenyl-1-propyl(2S)-1-(2-thienyl)glyoxyl-2-pyrrolidinecarboxylate, and apharmaceutically acceptable salt, hydrate, or a mixture thereof,wheresaid compound has an affinity for FKBP-type immunophilins wherein theimmunophilin exhibits rotamase activity and the compound inhibits therotamase activity of the immunophilin.